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Mutant construction for ychO, luxS and qseC genes present in an Avian Pathogenic Escherichia coli (APEC): in vivo and in vitro analysis

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Author(s):
Livia Pilatti Mendes Da Silva
Total Authors: 1
Document type: Master's Dissertation
Press: Campinas, SP.
Institution: Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia
Defense date:
Examining board members:
Wanderley Dias da Silveira; Vasco Ariston de Carvalho Azevedo; Marcelo Brocchi
Advisor: Wanderley Dias da Silveira
Abstract

Avian pathogenic Escherichia coli strains cause extraintestinal diseases in poultry, leading to substantial economic losses to the poultry industry worldwide. Virulence factors not yet known may play important roles in pathogenicity, which may be significant for measures control development of infectious processes of these strains. Invasins allow the pathogen to invade host cells, overcoming host defenses by interacting with specific cell surface receptors. Based on analysis of APEC SEPT362 strain genome detected the presence of a putative invasin gene homologous to ychO, not yet characterized, of E. coli str. K-12 substr. MG1655, and its effect on pathogenicity was studied. In this work, the SEPT362 strain deficient for ychO reduced the mortality rate in chicks, the adherence to chicken embryo fibroblast (CEF) cells, the invasion of chicken embryo cells (CEC-32), the survival in poultry macrophages (HD11), the motility and biofilm formation. These results suggest, for the first time, that ychO gene encodes for an invasin and plays a role in APEC SEPT362 pathogenicity. In addition to the many virulence factors, the study of different types of extracellular signals that mediate communication between bacteria and regulate gene expression opens up great possibilities to study pathogenic mechanisms and possible ways to interfere in it, transforming pathogenic organisms into non-virulent organisms. Quorum sensing (QS) is a mechanism dependent on the bacterial cell density that enables them to act as multicellular complexes, increasing their chances of survival in the environment. The QS system autoinducer 2 (AI- 2) is a universal communication system of bacteria mediated by AI- 2 produced by the action of the enzyme LuxS. The QS system autoinducer 3 (AI- 3) is a system that can also mediate communication between realms, being regulated by kinase QseC. In this context, mutants for the luxS and qseC genes were built in SEPT362 strain. The strain deficient for luxS showed a significant reduction in CEF cell adhesion due to a role in the regulation of gene type 1 fimbriae. The deficient strain for qseC had a lower rate of mortality in chicks and a significant reduction in cell adhesion to CEF, due to a lack of type 1 fimbriae in the mutant (AU)

FAPESP's process: 12/04391-1 - Mutation in AI-2 Quorum Sensing system and analyses of gene expression in an Avian Pathogenic Escherichia coli (APEC) strain
Grantee:Livia Pilatti Mendes da Silva
Support Opportunities: Scholarships in Brazil - Master